通过超分辨率成像对单个急性髓系白血病细胞的白细胞介素 3 受体α亚基进行定量分析。

Quantitative Profiling of Alpha-Subunit of IL-3 Receptor on Single Acute Myeloid Leukemia Cells by Super-Resolution Imaging.

机构信息

Institute for Synthetic Biology Research, Shenzhen Institutes of Advanced, Technology, Chinese Academy of Sciences, Shenzhen 518055, P.R. China.

National Laboratory of Biomacromolecules, CAS center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

出版信息

Asian Pac J Cancer Prev. 2023 Jan 1;24(1):185-194. doi: 10.31557/APJCP.2023.24.1.185.

DOI:10.31557/APJCP.2023.24.1.185

PMID:36708567

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10152859/

Abstract

BACKGROUND

Quantitative profiling of specific cell surface markers is a new approach in characterization of tumor heterogeneity and single cell biology. The current tools have dearth in detection and quantification of receptor proteins on single cells.

METHODS

we focused on our newly developed protocol to determine the distribution pattern and density of cell surface markers on single acute myeloid leukemia cells. Cell surface proteins were labeled with quantum dots (Qdots) followed by super resolution Structured Illumination Microscopy (SIM) imaging to imprisonment the optical signals emitted by Qdots which were further analyzed by software imaris to do three dimensional (3D) structure reconstruction and digital simulation. Furthermore, MTT assays and flow cytometry was performed to establish association between expression of cell surface markers and drug response.

RESULTS

In the present study, we found that the Molm13 and cytarabine-enriched Molm13 cells exhibit different densities of CD123, an alpha-subunit of interleukin-3 receptor, i.e. 0.92 and 1.73 per μm2 of cell surface respectively. Sub-populations of Molm13 cells expressing higher densities of CD123 on cells membranes showed resistance against cytarabine. Further study revealed that romidepsin sensitized and augmented the cytotoxicity of cytarabine in Molm13 and cytarabine-enriched Molm13 cells. Romidepsin increased the percentage of cell death-induced by cytarabine from 21.6 % to 28.6 % and 37.1 % to 57.2 % in Molm13 and cytarabine-enriched Molm13 cells respectively.

CONCLUSION

Altogether, the study suggests that Molm13 cells have sub-populations with differential expression of CD123+ phenotype. Romidepsin sensitizes and augments the effect of cytarabine in Molm13 and cytarabine-enriched Molm13 cells.

摘要

背景

定量分析特定细胞表面标志物是肿瘤异质性和单细胞生物学研究的新方法。目前的工具在检测和量化单细胞受体蛋白方面存在不足。

方法

我们专注于我们新开发的方案，以确定单个急性髓系白血病细胞表面标志物的分布模式和密度。细胞表面蛋白用量子点（Qdots）标记，然后用超分辨率结构光照明显微镜（SIM）成像来捕获 Qdots 发出的光学信号，进一步用软件 imaris 进行分析，进行三维（3D）结构重建和数字模拟。此外，进行 MTT 测定和流式细胞术，以建立细胞表面标志物表达与药物反应之间的关系。

结果

在本研究中，我们发现 Molm13 和阿糖胞苷富集的 Molm13 细胞表面的白细胞介素-3 受体α亚单位 CD123 的密度不同，分别为 0.92 和 1.73 个/μm2。细胞膜上表达更高密度 CD123 的 Molm13 细胞亚群对阿糖胞苷具有抗性。进一步的研究表明，罗米地辛增敏并增强了 Molm13 和阿糖胞苷富集的 Molm13 细胞中阿糖胞苷的细胞毒性。罗米地辛使阿糖胞苷诱导的细胞死亡百分比从 Molm13 细胞中的 21.6%增加到 28.6%，从阿糖胞苷富集的 Molm13 细胞中的 37.1%增加到 57.2%。